Recombinant CCN1 protein (wild-type, DM and D125A mutants) were produced in insect cells using a baculovirus system and purified as described (8 (link)). Immuno-blotting employed rabbit polyclonal anti-phospho-p53 (Ser15) and mAb against phospho-p38 MAPK (Thr180/Tyr182, D3F9), lamin A/C (4C11), and cytochrome c (D18C7) were from Cell Signaling Technology. Rabbit anti-EGFR (EP38Y) mAb was from AbCam. Rat anti-integrin α6 mAb (GoH3) was from Beckman Coulter.
Protocol detail
Purification and Analysis of Recombinant CCN1
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Baculovirus
Ccn1
Ccn1 protein
Cells
Cytochrome c
Egfr
Insect
Integrin
Lamin a c
P38 mapk
Rabbit
Recombinant protein
Corresponding Organization :
Other organizations : University of Illinois Chicago
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Variable analysis
independent variables
- Recombinant CCN1 protein (wild-type, DM and D125A mutants)
- Phosphorylation of p53 (Ser15)
- Phosphorylation of p38 MAPK (Thr180/Tyr182)
- Levels of lamin A/C
- Levels of cytochrome c
- Levels of EGFR
- Levels of integrin α6
- Experimental conditions for producing and purifying the recombinant CCN1 protein (wild-type, DM and D125A mutants) in insect cells using a baculovirus system as described in the referenced publication (8)
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